DEVELOPMENT OF PRIMER SETS DESIGNED FOR USE WITH THE PCR TO AMPLIFY CONSERVED GENES FROM FILAMENTOUS ASCOMYCETES

We constructed nine sets of oligonucleotide primers on the basis of th e results of DNA hybridization of cloned genes from Neurospora crassa and Aspergillus nidulans to the genomes of select filamentous ascomyce tes and deuteromycetes (with filamentous ascomycete affiliations). Nin e sets of primers were designed to amplify segments of DNA that span o ne or more introns in conserved genes. PCR DNA amplification with the nine primer sets with genomic DNA from ascomycetes, deuteromycetes, ba sidiomycetes, and plants revealed that five of the primer sets amplifi ed a product only from DNA of the filamentous ascomycetes and deuterom ycetes. The five primer sets were constructed from the N. crassa genes for histone 3, histone 4, beta-tubulin, and the plasma membrane ATPas e. With these five primer sets, polymorphisms were observed in both th e size of and restriction enzyme sites in the amplified products from the filamentous ascomycetes. The primer sets described here may provid e useful tools for phylogenetic studies and genome analyses in filamen tous ascomycetes and deuteromycetes (with ascomycete affiliations), as well as for the rapid differentiation of fungal species by PCR.